刷到了一个单细胞文章,是关于 amyotrophic lateral sclerosis (ALS), 是一种 neurodegenerative diseases ,我之前分享过:两个神经退行性疾病的单细胞核转录组队列的细胞亚群及其标记基因的比较,我看到这个文章的时候它仍然是在预印本:bioRxiv preprint doi: https://doi.org/10.1101/2022.03.25.485808; this version posted March 28, 2022.
文章标题是:《The Human Motoneuron Expression Signature is Defined by ALS-Related Genes》
取样和单细胞策略是:Lumbar spinal cord tissue was obtained from seven subjects (male and female, ~50-80 years old) and processed for single nucleus RNA sequencing.
降维聚类分群后,算法层面是65个亚群,生物学归类主要是是:
- oligodendrocytes and their precursors and progenitors,
- meningeal cells,
- astrocytes,
- endothelial
- pericyte cells,
- microglia,
- neurons;
- Schwann cells,
其中 neurons 还可以细分为:
- motoneurons (MN),
- excitatory dorsal neurons (ExDorsal),
- inhibitory dorsal neurons (InhDorsal),
- excitatory mid and ventral neurons (ExMV),
- inhibitory or mixed mid and ventral neurons (InhMV).
如下所示:
降维聚类分群
而且文章也给出来了各个单细胞亚群的特异性基因列表:
各个单细胞亚群的特异性基因列表
其实在:两个神经退行性疾病的单细胞核转录组队列的细胞亚群及其标记基因的比较,我列出来了一些基因:
代码语言:javascript复制astrocytes = c("AQP4", "ADGRV1", "GPC5", "RYR3")
endothelial = c("CLDN5", "ABCB1", "EBF1")
excitatory = c("CAMK2A", "CBLN2", "LDB2")
inhibitory = c("GAD1", "LHFPL3", "PCDH15")
microglia = c("C3", "LRMDA", "DOCK8")
oligodendrocytes = c("MBP", "PLP1", "ST18")
OPC='Tnr,Igsf21,Neu4,Gpr17'
Ependymal='Cfap126,Fam183b,Tmem212,pifo,Tekt1,Dnah12'
pericyte=c( 'DCN', 'LUM', 'GSN' ,'FGF7','MME', 'ACTA2','RGS5')
# 下面是4种神经细胞
# excitatory (SLC17A6),
# inhibitory (GAD2),
# GABAergic (GAD2/GRIK1),
# dopaminergic neurons(TH)
可以看到,在这个文章里面也是如此,蛮特意的。
这样的单细胞基础认知,也可以看基础10讲:
- 01. 上游分析流程
- 02.课题多少个样品,测序数据量如何
- 03. 过滤不合格细胞和基因(数据质控很重要)
- 04. 过滤线粒体核糖体基因
- 05. 去除细胞效应和基因效应
- 06.单细胞转录组数据的降维聚类分群
- 07.单细胞转录组数据处理之细胞亚群注释
- 08.把拿到的亚群进行更细致的分群
- 09.单细胞转录组数据处理之细胞亚群比例比较
最基础的往往是降维聚类分群,参考前面的例子:人人都能学会的单细胞聚类分群注释